Multiplex lateral flow test strip for detection of carbapenemase genes using barcoded tetrahedral DNA capture probe-based biosensing interface

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2023-08-22 DOI:10.1007/s00604-023-05903-y

Yongjie Xu, Jie Luo, Wei Lai, Jingjing Da, Bin Yang, Xiangrong Luo, Lin Zhan, Ying Fei, Lin Liu, Yan Zha

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引用次数: 1

Abstract

Carbapenem-resistant Enterobacterales pose significant global health challenges due to their rapid spread and ability to hydrolyse various beta-lactam antibiotics. Rapid tests for these carbapenemase genes are crucial to ensure appropriate prescription administration and infection control. In this study, we developed a rapid visual nanodiagnostic platform for multiplexed detection of carbapenemase genes using a lateral flow strip. The nanodiagnostic strip was designed with separate barcoded DNA tetrahedrons for the blaKPC and blaNDM genes. These tetrahedrons were distributed on a nitrocellulose membrane at two different test lines as capture probes. When tested against a panel of carbapenemase genes, the tetrahedral probes captured single-stranded amplicons of asymmetric PCR via strand hybridisation. The amplicons acted as bridging elements, binding the DNA-modified gold nanoparticles to the test line of the strip, resulting in clear visual readouts specific to the blaKPC and blaNDM genes. By employing barcoded tetrahedrons and asymmetric PCR in conjunction with the lateral flow strip, a single diagnostic test enabled the detection of multiple carbapenemase genes. The test yielded results as low as 0.12 fM for blaKPC and 0.05 fM for blaNDM within 75 min. Furthermore, the strip effectively identified specific carbapenemase genes in clinical isolates using real-time PCR, antibody-based lateral flow systems for carbapenemase detection, and carbapenemase phenotype experiments. Thus, the strip develop has a high potential for testing blaKPC and blaNDM genes in practice.

Graphical Abstract

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使用基于条形码四面体DNA捕获探针的生物传感界面检测碳青霉烯酶基因的多重横向流动测试条。

碳青霉烯类耐药肠杆菌由于其快速传播和水解各种β-内酰胺类抗生素的能力，对全球健康构成了重大挑战。对这些碳青霉烯酶基因的快速检测对于确保适当的处方给药和感染控制至关重要。在这项研究中，我们开发了一个快速视觉纳米诊断平台，用于使用侧流条对碳青霉烯酶基因进行多重检测。纳米诊断条是用blaKPC和blaNDM基因的单独条形码DNA四面体设计的。这些四面体分布在硝化纤维膜上的两个不同的测试线作为捕获探针。当针对碳青霉烯酶基因组进行测试时，四面体探针通过链杂交捕获不对称PCR的单链扩增子。扩增子充当桥接元件，将DNA修饰的金纳米颗粒结合到条带的测试线上，从而产生针对blaKPC和blaNDM基因的清晰视觉读数。通过将条形码四面体和不对称PCR与侧流条结合使用，单个诊断测试能够检测多个碳青霉烯酶基因。该测试在75分钟内产生了blaKPC低至0.12 fM和blaNDM低至0.05 fM的结果。此外，该条带使用实时PCR、用于碳青霉烯酶检测的基于抗体的侧流系统和碳青霉烯蛋白酶表型实验，有效地鉴定了临床分离株中的特异性碳青霉烯酶类基因。因此，开发的条带在实践中检测blaKPC和blaNDM基因具有很高的潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.